• Korean Journal of Microbiology
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• v.16 no.1
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• pp.30-40
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• 1978

Detergent soluble fractions were obtained from T. ferrooxidans ATCC 13598 and the T. thiooxidans ATCC 8085 which were treated with 3% of Tween 20. The detergent soluble antigen(crude antigen) fractions of the T.ferrooxidans and the T.thiooxidans were subjected to hydroxyapatite. In the case of T.thiooxidans, further purification was carried out on the DEAE-cellulose column chromatography. The antigen fractions, such as the hydroxyapatite peak-1(Tf, HA-1) and peak-2 from T.ferrooxidans(Tf. HA-2) and hydroxyapatite peak-1(Tt, HA-1), DEAE-cellulose peak-1(Tt, DP-1) and peak-2(Tt, DP-2) from T. thiooxidans wre compared each other with the homologous and the heterologous and the heterologous antisera against to the Thiobacillus species. The hydroxyapatite peak-2 fraction from the T.ferrooxidans(Tf, HA-2) and DEAE-cellulose peak-2 fraction from the T.thiooxidans(Tt, DP-2) were represented the type-specific immuno-reactivities between the T.ferrooxidans and the T.thiooxidans on the several sets of double gel diffusioin analysis. The type-specific antigen fractions from both of the baceteria were mainly composed of protein with entierly different electrophoretic mobility on the SDS-polyacrylamide gel electrophoresis. However, the PAS positive banding patterns on the electrophorogram showed wide range of common antigenic properties in the T. ferrooxidans and the T.thiooxidans, respectively.

• Journal of Life Science
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• v.18 no.4
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• pp.494-502
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• 2008

The anaphylaxis shock reaction on the whole cells of H. pylori exhibited a symptom of slight illness for the first and second medication of causing antigen at an antigen concentration of WC (H) $60\;{\mu}g/100\;{\mu}l$ for WC (H) and no anaphylaxis shock symptom was observed at an antigen concentration of $20\;{\mu}g/100\;{\mu}l$ for WC (L). In the case of anaphylaxis shock reaction on the crude urease, no symptom was observed at an antigen concentration of $20\;{\mu}g/100\;{\mu}l$ for both urease (L) and urease (H). In the heterologous passive cutaneous anaphylaxis (PCA) test using a guinea pig-rat, no positive reaction was detected in all the medication groups of WC (H), WC (L), urease (H) and urease (L). In the skin sensitization test, it was observed that the best antigen concentration not causing skin disorder at each of $80\;{\mu}g/100\;{\mu}l$, $40\;{\mu}g/100\;{\mu}l$, $20\;{\mu}g/100\;{\mu}l$, and $20\;{\mu}g/100\;{\mu}l$ was $40\;{\mu}g/100\;{\mu}l$.

• The Journal of Korean Society of Virology
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• v.27 no.2
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• pp.161-168
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• 1997

The present study was designed to estimate the seroprevalence of NLVs among diarrheagenic children and in healthy adults in Seoul and its vicinity with the use of an EIA and an Western blot (WB) based on recombinant Norwalk virus capsid protein (rNV) and crude virus preparations as antigen. Seroconversion was observed in 34 (83%) of 41 tested using the EIA and in 21 (54%) of 39 using the WB, suggesting that the NLVs with epitopes common to rNV are prevalent in Seoul area. Diarrheal children who were known to have been infected with several other strains of the NLVs showed no significant antibody response to the rNV. Infection with rNV occurred earlier in life: primary infections with rNV were common before the age of 6 months and over 91 % of children had evidence of infection by that age by the EIA. Since the amount of the NLV antigens available for seroepidemiologic surveys is limited, we tried to detect NLV antibody by using crude virus preparations as antigen. One crude virus preparation of a child whose stool yielded genetically distinct NLV revealed the presence of the plural number of bands upon SDS-PAGE, but precipitated only one band (62 kDa) after the WB with a serum (collected 10 days after the onset of symptoms) of another diarrheal child. The WB assay we present in this report revealed that the NLVs are prevalent among Korean population and that the sera contained antibody to a single major structural protein, with molecular sizes of 58 to 62 kDa, compatible with the sizes reported for the Norwalk virus and Snow Mountain agent proteins, respectively. When the results of the WB were compared with those obtained by the EIA, the EIA antibody assay was sensitive enough to detect an antibody rise of as much as 4096-fold but not as specific as the WB. The WB assay presented in this paper will provide a powerful tool to elucidate not only antigenic structures of the NL Vs but also seroepidemiology of the NLV infection. The availability of an unlimited source of antigen will enable a large scale serologic studies that will greatly increase our understanding of the role of NLVs in human enteric illness.

• Parasites, Hosts and Diseases
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• v.44 no.1 s.137
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• pp.43-48
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• 2006

Experimental murine models with high, intermediate and low levels of genetically based susceptibility to Leishmania major infection reproduce almost entire spectrum of clinical manifestations of the human disease. There are increasing non-comparative studies on immune responses against isolated antigens of L. major in different murine strains. The aim of the present study was to find out whether there is an antigen that can induce protective immune response in resistant and susceptible murine strains. To do that, crude antigenic extract of procyclic and metacyclic promastigotes of L. major was prepared and subjected to SDS-PAGE electrophoresis. Western-blotting was used to search for antigen(s) capable of raising high antibody level of IgG2a versus IgG1 in the sera of both infected resistant and susceptible strains. Two novel antigens from metacyclic promastigotes of L. major (140 and 152 kDa) were potentially able to induce specific dominant IgG2a responses in BALB/c and C57BU6 mice. The 2 antigens also reacted with IgG antibody of cutaneous leishmaniasis patients. We confirm that 140 and 152 kDa proteins of L. major promastigotes are inducing IgG production in mice and humans.

• Parasites, Hosts and Diseases
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• v.27 no.1
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• pp.9-14
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• 1989

Enzyme-linked immunoelectrotransfer blot (EITB) using crude worm antigen of adult Paragonimus westermani was performed for human patients sera to identify the species-specific components. Crude antigen was obtained by homogenizing and centrifuging 24-week old adult worms at 10,000 rpm for 60 minutes in phosphate buffered saline (PBS, PH 7.2) containing: Phenyl methyl sulfonyl auoride (PMSF). Gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) was performed and blotted electrophoretically onto a sheet of nitrocellulose paper. The sheet was cut into strips and exposed to sera diluted 1 : 200 with PBS. SDS-PAGE showed 26 protein bands ranging 229 to 10 kDa. Of them 229, 91, 60, 50, 35∼31, 27, 25, 21, 17, 11 and 10 kDa components showed positive reaction with serum antibody of patients with p. westermani. Sera of patients infected with Clcnorchis sinensis reacted with 35∼31, 19, and 11 kDa bands. Human sera from cysticercosis and diphyllobothriasis cases showed non-specific cross reactions with 229, 35∼31, 27, 25 and 17kDa bands. Protein bands of 91, 60, 21 and 10kDa showed strong positive reaction without cross reactions with sera from other helminthic infections.

• Journal of agricultural medicine and community health
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• v.16 no.1
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• pp.61-69
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• 1991

Serodiagnosis of Clonorchis sinensis infections will probably be a first choice tool for screening of clonorchiasis in a future because of increasing difficulties in collection and examination of stools. The sensitive test such as ELISA can he used effectively. However there are some limitations in serological diagnosis for the detection of serum antibody. One of the major problems is the non-specificity of the antigens which produce cross reaction with other helminthic infection sera. To solve this problem. many investigators have tried to purify the antigens used. In this study, we determined the antigenic profile of the crude saline extract antigen of C. sinensis at early developmental stage based on SDS-PAGE and immunoblotting techniques for the purpose of understanding the nature of C. sinensis worm antigen The following results were obtained : 1) The SDS-PAGE showed many protein hands ranging from 10Kd to 91Kd relative molecular weight. Among them, 66, 46, 40, 33, 27, 24, 16, 14 and 10Kd bands were observed as a principle bands. The protein components of C. sinensis changed chronologically during their early developmental period. 44Kd band was stained unclearly in antigen of 2 weeks worm, but changed to concentrated state in antigen of 5 weeks worm. 35Kd band was found in antigen of 2 weeks worm, however this band was disappeared in antigen of 5 weeks worm. 22Kd band also lost its staining property gradually. 2) In spite of differences in antigenic profile, there was no differences in the data obtained by microplate ELISA using each antigen preparation. Absorbance value began to rise in between 2 to 3 weeks after infection. 3) By EITB. serum antibody recognized major protein bands with molecular weight of 91, 85, 63, 46, 40, 33, 24, 14 and 10Kd hand respectively. Among them 66, 33, 17 and 14Kd bands were observed as non-specific band because they reacted even in normal control sera. Generally, gradual increase of positive reactions were observed as the infection period of C. sinensis was prolonged. In other hand, the reaction of 10Kd hand did not occurred when 26th week sera was tested. 4) The positive reactions using antigens of 2 weeks worm, especially on 40 and 24Kd bands, were most strong and sharply demarcated compared to those of 3~5 weeks worm antigen.

• Parasites, Hosts and Diseases
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• v.21 no.2
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• pp.270-280
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• 1983

Agar-gel Diffusion test (AGD), Counterimmunoelectrophoresis(CIEP) and Enzyme-Linked Immunosorbent Assay(ELISA) were examined with the sera of skin test positives for paragonimiasis. The crude antigen (Paragcnimus whole worm extracts: protein concentration, 7.56mg/m1) and human sera were used in AGD and CIEP. And in ELISA test, diluted antigen with 1:40, 000 of crude antigen and diluted sera with 1:100, 1:200 were used in the test. The positive identical ratio between AGD and CIEP reactions is 985 and negative identical ratio is 100%. One or three precipitin bands are observed in AGD. One to seven precipitin bands are also revealed in CIEP. Especially, deeply stained bands are observed in CIEP than those of AGD. The positive identical ratios between AGD and ELISA tests are 96% in 1:100 diluted sera, and 94% in 1:200 diluted sera. But the negative identical ratios between AGD and ELISA tests are 97% and 99% respectively in 1:100 and 1:200 diluted sera. The positive identical ratios between CIEP and ELIEA tests are 98% and 96% respectively in 1:100 and 1:200 diluted sera, but also 97% and 99% in 1:100 and 1:200. Control sera, such as clonorchiasis, amoebiasis and toxoplasmcsis, revealed all negatives with Paragonimus antigen in AGD, CIEP and ELISA tests. By above results, ELISA was mcst sensitive, next CIEP and AGD, But AGD test apprars to be more useful when used to crude antigen without cross rfacticn with other parasitic infections. CIEP test is basically equal in terms of precipitin reaction, but CIEP is able to be detected more sensitively and rapidly though less simple in handiwork than AGD. Consequently, three methods for inlmunological tests of paragonimiasis have gccd correlations with one another. Also, each of these has both merits and demerits in iymunolcgical test for paragonimiasis. But the ELISA test was proved to be the most sensitive and convenient tool for mass screening test, especially in cacti of using purified antigen.

• Parasites, Hosts and Diseases
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• v.42 no.4
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• pp.169-174
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• 2004

This study was carried out to find out specific proteins from different organs of Clonorchis sinensis. Crude extract, organ-specific and excretory-secretory (ES) proteins were analyzed by immunoblot with infected human sera. The bands of 7- and 17 -kDa were main component of intestinal fluid and ES protein and commonly found in all organ-specific proteins. The 17-kDa protein was observed from ES antigen, intestinal fluid, eggs and sperms, 26- and 28-kDa proteins were from the uterus, vitellaria, and ovary, and 34-, 37-, 43- and 50-kDa proteins were mainly from the testis and sperms. Serum of mice immunized with sperms reacted to the 50-kDa protein by immunoblotting and immunohistochemical staining showed a positive reaction at the seminal receptacle and seminiferous tubule. The present results show that the 7-kDa protein is a common antigen of every part or organ of C. sinensis, but different organs express their specific antigenic protein bands.

• Parasites, Hosts and Diseases
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• v.51 no.6
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• pp.637-644
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• 2013

This study aimed to investigate the antibody responses in mice immunized with Gnathostoma spinigerum crude antigen (GsAg) incorporated with the combined adjuvant, a synthetic oligonucleotide containing unmethylated CpG motif (CpG ODN 1826) and a stable water in oil emulsion (Montanide ISA720). Mice immunized with GsAg and combined adjuvant produced all antibody classes and subclasses to GsAg except IgA. IgG2a/2b/3 but not IgG1 subclasses were enhanced by immunization with CpG ODN 1826 when compared with the control groups immunized with non-CpG ODN and Montanide ISA or only with Montanide ISA, suggesting a biased induction of a Th1-type response by CpG ODN. After challenge infection with live G. spinigerum larvae, the levels of IgG2a/2b/3 antibody subclasses decreased immediately and continuously, while the IgG1 subclass remained at high levels. This also corresponded to a continuous decrease of the IgG2a/IgG1 ratio after infection. Only IgM and IgG1 antibodies, but not IgG2a/2b/3, were significantly produced in adjuvant control groups after infection. These findings suggest that G. spinigerum infection potently induces a Th2-type biased response.

• Journal of Ginseng Research
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• v.30 no.3
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• pp.117-127
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• 2006

Ginseng is a medicinal herb widely used in Asian countries. Dendritic cells(DCs) play a pivotal role in the initiation of T cell-mediated immune responses, making them an attractive cellular adjuvant for use in cancer vaccines. In this study, we examined the effects of Red-ginseng(water extract, edible and fermented ethyl alcohol extract, crude saponin) on the DCs phenotypic and functional maturation. Immature DCs were cultured in the presence of GM-CSF and IL-4, and the generated immature DCs were stimulated by water extract, edible and fermented ethyl alcohol extract, crude saponin and LPS, respectively, for 24hours. The expression of surface co-stimulatory molecules, including MHC(major histocompatibility complex) class II, CD40, CD80 and CD86, was increased on DCs that were stimulated with crude saponin, but antigen-uptake capacity was decreased. The antigen-presenting capacity of Red-ginseng extracts-treated DCs as analyzed by allogeneic T cells proliferation and IL-2, $IFN-{\gamma}$ production was increased. Furthermore, $CD4^+$ and $CD8^+$ syngeneic T cell(OVA-specific) proliferation and $IFN-{\gamma}$ production was significantly increased. However, $CD4^+$ syngeneic T cell secreted higher levels of IL-2 in responding but not $CD8^+$ syngeneic T cell. These results indicate the immunomodulatory properties of Red-ginseng extracts, which might be therapeutically useful in the control of cancers and immunodeficient diseases through the up-regulation of DCs maturation.